Tylka Nematology Lab

Procedure:

• Wash cyst/sediment suspensions from beakers into 50 ml plastic centrifuge tubes.
• Adjust the volume of liquid in each tube to approximately the same level (to avoid excessive centrifuge vibration).
• Place the tubes, 2 by 2, into opposite positions in the centrifuge.
• Centrifuge the tubes at 2000 RPM for 4 minutes. After 4 minutes, turn the centrifuge off and let the rotor coast to a stop.
• Save the supernatant and pour into the original sample beaker.
• Wipe the rim of the centrifuge tube to collect cysts stuck to the tube wall, and rinse into beaker.
• Add sucrose solution (1362 g sucrose / 1L tap water) to tubes until each is about 2/3 full, and stir or agitate the tubes to re-suspend the sediments and cysts. After stirring, add more sucrose solution to level off paired tubes.
• Centrifuge at 2000 RPM for 2 minutes. After 2 minutes, turn the centrifuge off and let the rotor coast to a stop.
• For each sample, pour the supernatant through a #60 or #80 mesh sieve. In addition, pour the supernatant from the 1st centrifuge step through the sieve. Rinse thoroughly with tap water.
• Collect cysts by rinsing them from the sieve back into the original beaker using a water bottle.
• Wash the plug out of the bottom of the tubes, and clean and rinse the tubes.